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Real-time MBDi-RPA using methyl-CpG binding protein 2: A real-time detection method for simple and rapid estimation of CpG methylation status
Get PDFBackground
Analysis of CpG methylation is informative for cancer diagnosis. Previously, we developed a novel method to discriminate CpG methylation status in target DNA by blocking recombinase polymerase amplification (RPA), an isothermal DNA amplification technique, using methyl-CpG binding domain (MBD) protein 2 (MBD2). The method was named MBD protein interference-RPA (MBDi-RPA). In this study, MBDi-RPA was performed using methyl-CpG binding protein 2 (MeCP2), another MBD family protein, as the blocking agent.
Results
MBDi-RPA using MeCP2 detected low levels of CpG methylation, showing that it had higher sensitivity than MBDi-RPA using MBD2. We also developed real-time RPA, which enabled rapid analysis of DNA amplification without the need for laborious agarose gel electrophoresis and used
it in combination with MBDi-RPA. We termed this method real-time MBDi-RPA. The method using MeCP2 could determine the abundance ratio of CpG-methylated target DNA simply and rapidly, although highly sensitive detection was challenging.
Significance and Novelty
Real-time MBDi-RPA using MeCP2 could be potentially useful for estimating CpG methylation status in target DNA prior to more detailed analyses.弘前大学博士（医学）Author: Mina Ishidoya, Toshitsugu Fujita, Sadatomo Tasaka, Hodaka FujiiCitation: Analytica Chimica Acta, 1302:342486, 2024 May
Circular RNAs hsa_circ_0001438 and hsa_circ_0000417 are downregulated and upregulated, respectively, in hepatocellular carcinoma
Get PDFHepatocellular carcinoma (HCC) is the most predominant type of liver cancer and is frequently fatal. Alpha-fetoprotein, alpha-fetoprotein-L3, and protein induced by vitamin K absence or antagonist-II are used as biomarkers to diagnose HCC. However, these biomarkers are not highly specific, especially for early-stage HCC diagnosis; therefore, more specific biomarkers are needed. Recently, circular RNA (circRNA) biomarkers have been used to diagnose several intractable diseases. In this study, we sought to identify circRNA biomarkers for the specific diagnosis of HCC. To this end, we compared the expression levels of circRNAs in primary HCC and normal tissues using publicly available RNA-seq data. Our analysis revealed that the expression levels of eight circRNAs were altered in primary HCC tissues compared with normal tissues. To confirm our findings, we examined the expression levels of selected circRNAs in HCC cell lines and normal hepatocytes. The expression level of hsa_circ_0001438, a circRNA that was downregulated in primary HCC, was lower in poorly and well-differentiated HCC cell lines than in normal hepatocytes. By contrast, the expression level of hsa_circ_0000417, which was increased in primary HCC, was strongly upregulated in a well-differentiated HCC cell line compared with normal hepatocytes. Thus, hsa_circ_0001438 and hsa_circ_0000417 might be potential biomarkers for the specific diagnosis of HCC. The experimental strategy described here, using publicly available RNA-seq data, is a useful and cost-effective method of identifying circRNA biomarkers.弘前大学博士（医学）Author: Sachiko Imanishi, Shoko Nagata, Toshitsugu Fujita, Hodaka FujiiCitation: International Journal of Experimental Pathology 103(6):245-251, 2022 De
The effects of combined early oral health care and early mobilisation on the incidence of stroke-associated pneumonia
Get PDFObjective: To investigate the effects of combined early oral healthcare and early mobilisation on the incidence of stroke-associated pneumonia during hospitalisation of acute stroke patients.
Materials and methods: In this single-centre, non-blinded, before-and-after cohort study, patients received basic stroke rehabilitation by a multidisciplinary team within 72 h of symptom onset from July to September 2016 and from July to September 2018. Patients were divided into two groups: 1) patients who received combined early oral healthcare and early mobilisation (early intervention group) (n=107), and 2) patients who received usual care (control group) (n=107). The relationship between the stroke-associated pneumonia incidence and prognosis was examined.
Results: The early intervention group had a significantly lower incidence of stroke-associated pneumonia than the control group (0.93% vs. 7.48%; P=0.01). Moreover, the early intervention group had a significantly lower proportion of patients who died or required medical care because of recurrent pneumonia at discharge (0.93% vs. 5.6%; P=0.04). In contrast, there were no significant differences between the two groups regarding the Revised Hasegawa's Dementia Scale on day 14 (22.5 vs. 23; P=0.87), Functional Independence Measure on day 14 (112 vs. 116; P=0.06), and rate of total oral diet (Food Intake LEVEL Scale ≥7) at discharge (95.2% vs. 93.5%; P=0.55).
Conclusions: Combined early oral healthcare and early mobilisation by a multidisciplinary team significantly decreased the stroke-associated pneumonia incidence within 7 days and reduced the percentage of patients who died or required medical care because of recurrent pneumonia after stroke.弘前大学博士（医学）Author: Ayaka Fujita, Manabu Iwata, Joji Hagii, Norifumi Metoki, Hiroshi Shiroto, Eiichi TsudaCitation: Journal of Stroke and Cerebrovascular Diseases ;32(1):106893, 2023 Ja
Association between regional oxygen saturation and central venous saturation in pediatric patients undergoing cardiac surgery: A prospective observational study
Get PDF弘前大学博士（医学）掲載誌：Pediatric Anesthesia 2023 Nov;33(11):913-922., Epub 2023 Aug 3
Video vs. ultrasound pupillometry for detecting increased pupillary diameters due to nociceptive stimuli: A prospective observational study
Get PDF弘前大学博士（医学）掲載誌：Journal of Anesthesia, 2024 Apr;38(2):191-197., Epub 2024 Jan 8
Creation of Tsugaru Dialect Dictionary Database Based on Tsugaru Dialect Corpus and Proposal of Usage as Educational Tool
Get PDF弘前大学修士（人文社会科学
Factors associated with the effect of anamorelin on cachexia in patients with non-small cell lung cancer
Get PDFPURPOSE: Anamorelin, a ghrelin receptor agonist, is approved in Japan for the treatment of cachexia in patients with lung and gastrointestinal cancer. However, there is limited research on the usefulness of anamorelin in clinical settings.
METHODS: A total of 40 patients with non-small cell lung cancer and cachexia who were prescribed anamorelin between July 2021 and November 2022, were retrospectively assessed. A logistic regression analysis was used to analyze the association between background characteristics and early discontinuation of treatment with anamorelin （within 4 weeks）.
RESULTS: The median age was 67 years （range, 36-88）, and 65% of the patients were male. The early discontinuation group included 11 patients （27.5%）. An Eastern Cooperative Oncology Group Performance Status （ECOG-PS） score 2 （odds ratio, 7.85; 95% confidence interval, 1.43-43.21; P=0.018） was associated with early discontinuation. A total of 18/40 patients were able to continue anamorelin treatment for 12 weeks, and the mean change in body weight was +2.31 kg, which was a significant change from the weight recorded at baseline（ P=0.027）.
CONCLUSION: This study indicates that anamorelin is unlikely to be useful for patients with a poor general condition（ ECOG-PS score 2）.弘前大学博士（医学）Author: Yoshiko Ishioka, Hisashi Tanaka, Tomonori Makiguchi, Hiroaki Sakamoto, Toshihiro Shiratori, Kageaki Taima, Sadatomo TasakaCitation: Hirosaki Medical Journal 75：22-30，202
Impact of factor Xa inhibitors on outcomes of atrial tachyarrhythmia recurrence following catheter ablation for atrial fibrillation: comparison with warfarin
Get PDF弘前大学博士（医学）掲載誌：Hirosaki Med. J. 74：87-98，202